SITE-DIRECTED MUTAGENESIS BY DOUBLE POLYMERASE CHAIN-REACTION

被引：77

作者：

BARIK, S ^{[1
]}

机构：

[1] UNIV SO ALABAMA,COLL MED,DEPT BIOCHEM & MOLEC BIOL,MOBILE,AL 36688

来源：

MOLECULAR BIOTECHNOLOGY | 1995年 / 3卷 / 01期

关键词：

PCR; MUTAGENESIS; GENE CLONING;

D O I：

10.1007/BF02821329

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

A simple and versatile method of in vitro site-specific mutagenesis based on polymerase chain reaction (PCR) is described. The complete method required the use of three oligonucleotide primers and two PCRs. The product of the first PCR was used as one of the primers (megaprimer) in the second PCR. Essentially 100% of the final product incorporated the desired mutation. The various aspects of the procedure and its application is described in detail.

引用

页码：1 / 7

页数：7

共 10 条

[1] BARIK S, 1991, BIOTECHNIQUES, V10, P489
[2] BARIK S, 1991, BIOTECHNIQUES, V10, P490
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[J]. NUCLEIC ACIDS RESEARCH, 1988, 16 (20) : 9677 - 9686
[5] FINCKH U, 1991, BIOTECHNIQUES, V10, P35
[6] A GENERAL-METHOD OF INVITRO PREPARATION AND SPECIFIC MUTAGENESIS OF DNA FRAGMENTS - STUDY OF PROTEIN AND DNA INTERACTIONS
HIGUCHI, R
KRUMMEL, B
SAIKI, RK
[J]. NUCLEIC ACIDS RESEARCH, 1988, 16 (15) : 7351 - 7367
[7] MANIATIS T, 1982, MOL CLONING LABORATO, P113
[8] SARKAR G, 1990, BIOTECHNIQUES, V8, P404
[9] GENOMIC AMPLIFICATION WITH TRANSCRIPT SEQUENCING
STOFLET, ES
KOEBERL, DD
SARKAR, G
SOMMER, SS
[J]. SCIENCE, 1988, 239 (4839) : 491 - 494
[10] ZINTZ CB, 1991, BIOTECHNIQUES, V11, P158

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